PsbD | D2 protein of PSII

KLH-comjugated synthetic peptide derived from the C-terminal of known PsbD sequences including Arabidopsis thaliana P56761, Hordeum vulgare P11849, Chlamydomonas reinhardtii P06007, Synechococcus sp. PCC 7002 P20898

39.4 | 28-30 kDa

Application example

2 µg of total protein from (1) Arabidopsis thaliana leaf extracted with Agrisera Protein Extraction Buffer, PEB (AS08 300), (2) Hordeum vulgare leaf extracted with PEB, (3) Chlamydomonas reinhardtii total cell extracted with PEB, (4) Synechococcus sp. 7942 total cell extracted with PEB, (5) Anabaena sp. total cell extracted with PEB were separated on  4-12% NuPage (Invitrogen) LDS-PAGE and blotted 1h to PVDF. Blots were blocked immediately following transfer in 2% blocking reagent in 20 mM Tris, 137 mM sodium chloride pH 7.6 with 0.1% (v/v) Tween-20 (TBS-T) for 1h at room temperature with agitation. Blots were incubated in the primary antibody at a dilution of 1: 50 000 for 1h at room temperature with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at room temperature with agitation. Blots were incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, recommended secondary antibody AS09 602) diluted to 1:50 000 in 2% blocking solution for 1h at room temperature with agitation. The blots were washed as above and developed for 5 min with chemiluminescence detection reagent of extreme femtogram sensitivity, according the manufacturers instructions. Images of the blots were obtained using a CCD imager (FluorSMax, Bio-Rad) and Quantity One software (Bio-Rad). Exposure time was 3 seconds.

The peptide used to elicit this antibody has a perfect conservation across all full-length PsbD sequences from higher plants, lower plants, cyanobacteria and unicellular algae except:  minor substitutions in some Prochlorococcus & Dinoflagellate sequences. The antibody should still work against these taxa, but it has not been tested yet.  This antibody does not detect PsbA protein (D1).

There is a confirmed cross-reaction with TLA1 protein in Chlamydomonas reinhardtii.

For samples with a very low PSII content theremight be detection problems independent of the antibody. PSII proteins can vary in level depending upon liquid culture conditions. When the cells are in a stationary phase PSII content can drop to a very low level.

AS09 146S | PsbD | D2 protein of PSII protein standard for western blot quantitation and as a positive control
AS06 146PRE | PsbD | D2 protein of PSII, pre-immune serum, control for immunolocalization
AS05 084 | Anti-PsbA (D1) protein of PSII, C-terminal, rabbit antibodies
collection of antibodies to PSII proteins

D2 protein (PsbD) forms the reaction core of PSII (Photosystem II) as a heterodimer with the D1 protein (PsbA). PsbD is homologous to the D1 protein, with slightly higher molecular mass of about 39.5 kDa. Accumulation of D2 protein is an important step in the assemply of the PSII reaction centre complex.